猪的脂肪和骨髓来源间充质干细胞培养特性比较  

Comparison of culture characteristics between swine bone marrow and adipose tissue-derived mesenchymal stem cells

作　　者：尹巧香[1] 王恒[2] 裴志勇[3] 赵玉生[4] 

机构地区：[1]中国人民解放军空军总医院干部病房心内科,北京100142 [2]中国人民解放军空军总医院神经内科,北京100142 [3]中国人民解放军北京军区总医院干部病房一科,北京100070 [4]中国人民解放军总医院老年心血管病研究所,北京100853

出　　处：《生物医学工程与临床》2013年第6期523-527,共5页Biomedical Engineering and Clinical Medicine

基　　金："十五"国家"863计划"基金资助项目(2006AA02A105)

摘　　要：目的明确小型猪的脂肪来源间充质干细胞（A_r_MSCs）和猪的骨髓来源间充质干细胞（BM—MSCsl体外培养特性的异同。方法广西巴马小型猪，雌雄不限，猪龄4～6个月，体质量20～30kg。AT-MSCs来源于小型猪腹股沟皮下组织．BM—MSCs来源于小型猪的骨髓组织。培养AT-MSCs和BM—MSCs并观察它们的细胞形态。流式细胞仪检测Arr_MSCs和BM—MSCs的表面标志物（CD29、CD34、CD45、CD90）。分别观察Arr-MSCs和BM—MSCs的细胞生长分化能力；实时聚合酶链反应（PCR）检测基因表达。结果流式细胞仪检测结果表明，A．r_MSCs和BM—MSCs均表达CD29[分别为（99．06±0．30）％、（99．94±0．05）％]、CD90[分别为（97．404-0．40）％、（97．43±1J29）％1阳性，CD34、CD45阴性。AT-MSCs传代需培养5～7d，而BM—MSCs需培养7～10d。与BM—MSCs比较，AT—MSCs具有更强的生长分化能力。实时PCR检测基因表达结果显示．AT—MSCs和BM—MSCs均能分化心肌特异标志物a—skeletalactin和Troponin—I．二者差异无统计学意义．表明AT—MSCs和BM—MSCs均具备多项分化潜能．结论AT—MSCs是小型猪干细胞移植治疗的理想选择．Objective To investigate the differences of ex vivo culture characteristics between swine adipose tissue-derived mesenchymal stem cells （AT-MSCs） and bone marrow-derived mesenchymal stem cells（BM-MSCs）. Methods Chinese Bama swine with body weight of 20 - 30 kg were enrolled. AT-MSCs were harvested from inguinal subcutaneous fat tissue and BM- MSCs were isolated from bone marrow. AT-MSCs and BM-MSCs were cultured and the morphology of cells was observed. The surface antigens（CD29, CD34, CD45 and CD90） of AT- MSCs and BM-MSCs were examined by flow cytometry. The growth and proliferation, as well as gene expression by the real-time polymerase chain reaction of AT-MSCs and BM-MSCs were observed. Results Both AT-MSCs and BM-MSCs were determined as stem cells by flow cytometry, which showed positive expression of CD29[（99.06 ± 0.30） % vs （99.94 ± 0.05） %] and CD90[（97.40 ± 0.40） % vs （97.43 ± 1.29） %], and negative expression of CD34 and CD45. AT-MSCs could passage after cultured for about 5 - 7 days, but BM-MSCs needed about 7 - 10 days. Compared with BM-MSCs, AT-MSCs had more powerful ability of growth and proliferation. Quantitative real-time polymerase chain reaction revealed that there was no significant difference in expression of a-skeletal aetin and Troponin-I between BM-MSCs and AT- MSCs. BM-MSCs and AT-MSCs had multiple differentiation potential. Conclusion It is demonstrated that AT-MSCs are good choice for swine stem cell transplantation research.

关 键 词：猪 间充质干细胞 脂肪组织 骨髓 培养特性 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]